Anti-LAMP1 antibody - Lysosome Marker images

ab24170 staining Lamp-1 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab24170 at 1μg/ml and ab7291 (staining Tubulin) at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI, which was added to the secondary antibody mixture.

NOTE: The filamentous pattern observed only occurs when cell are fixed with 100% methanol. We are aware that the staining pattern can change if the cells are fixed with 4%PFA or an alternative fixative. LAMP-1 is a lysosome marker but also plays a role in cellular adhesion. Therefore, lysosomal and filamentous patterns for LAMP-1 can be possible.

IHC-P image of LAMP1 staining on human Cortex sections using ab24170 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retreival using citric acid. The sections were then permeabilized using 0.05% Tween-20 and blocking was performed using 3% BSA for 1 hour at 21°C. The primary antibody ab24170 was diluted using 3% BSA with 0.05% Tween-20 in PBS and incubated with the sections for 18 hours at 4°C. The secondary antibody used was Goat polyclonal to rabbit IgG conjugated to biotin (1:500)

ab24170 staining LAMP1 in human fibrosarcoma cells by Immunocytochemistry/ Immunofluorescence. The cells were PFA fixed, permeabilised in 0.1% Triton X-100 and incubated with the primary antibody at 1µg/ml for 1 hour at 20°C. The secondary antibody used was a donkey anti-rabbit (H+L) IgG conjugated to Alexa Fluor® 555 used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).

ICC/IF image of ab24170 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24170, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.